Exploiting the Salivary Arsenal to Inhibit Diarrheal Disease

利用唾液库抑制腹泻病

基本信息

批准号：
10357830
负责人：
ESTHER BULLITT
金额：
$ 20.44万
依托单位：
BOSTON UNIVERSITY MEDICAL CAMPUS
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-02-22 至 2025-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10357830
关键词：
Adherence Adhesives Anti-Bacterial Agents Antibiotic Resistance Antibiotics Bacteria Bacterial Adhesion Bacterial Infections Binding Biological Models Cell Culture Techniques Cell Wall Cells Cessation of life Child Cryoelectron Microscopy Data Development Diarrhea Disease Drug Design Effectiveness Enterobacteriaceae Enzymes Filament Functional disorder Gastrointestinal Diseases Gastrointestinal tract structure Goals Health Human Immune Innate Immune System Intestines Natural Immunity Nucleic Acids Organoids Oropharyngeal Outcomes Research Pathogenicity Peptides Periodontal Diseases Pilum Property Proteins Proteome Research Research Personnel Resolution Respiratory Disease Role Route Saliva Salivary Structure Surface Testing Therapeutic Therapeutic Agents Virulence Factors Vulnerable Populations antimicrobial antimicrobial peptide base combat design diarrheal disease efficacy testing enterotoxigenic Escherichia coli fighting histatin 5 inhibitor innovation novel novel strategies novel therapeutic intervention oral infection oral pathogen pathogen peptide drug prevent prophylactic respiratory respiratory pathogen therapeutic development

项目摘要

The ability of salivary peptides to inhibit diarrhea-causing bacteria from binding to the gut is newly discovered, and not well characterized. The innovative long-term goal is to realize the full potential of salivary components as both prophylactic and therapeutic agents against gastrointestinal and respiratory diseases. The objectives of this proposal are to define the efficacy of specific salivary components as pathogen inhibitors, and to determine the structure and mechanism by which one these peptides, Histatin-5, binds to pili that are external filaments on enterotoxigenic Escherichia coli (ETEC). This pilus/peptide interaction provides the dysfunction that inhibits bacterial binding, and knowing the mechanism of this dysfunction will lead to novel therapeutic approaches against ETEC and other pathogens. The central hypothesis is that enhanced utilization of the innate immune system to combat disease can be achieved through therapeutics developed from components of saliva. The rationale for this proposal is that completion of this research provides a path forward for utilizing salivary components to fight diarrheal diseases. In addition, determination of the mechanism of Histatin-5's action will permit creation of therapeutics with even greater efficacy. To achieve our goals, we will pursue the following two specific aims: 1. Define the capacity of selected salivary peptides to inhibit bacterial binding of enterotoxigenic Escherichia coli (ETEC) to target cells; 2. Determine the mechanism by which the salivary peptide, Histatin-5, inhibits bacterial binding via pili, an essential virulence factor of enterotoxigenic Escherichia coli (ETEC). These aims will be achieved using 1) bacterial adhesion studies on cell cultures and primary human intestinal cultures (“organoids”) and 2) structure determination at high resolution using electron cryomicroscopy and cryotomography (cryo-EM and cryo-ET). The proposed research is significant, because it will open a new avenue for development of therapeutics against diarrhea-causing bacteria. Unlike traditional antibiotics that broadly target enzymes involved in nucleic acid, protein, and cell wall synthesis, here, we explore a novel aspect of the host's innate immune system: the ability of salivary components to inhibit bacterial adherence to the host. The expected outcome of this research is a list of salivary components that can be exploited as therapeutics against diarrhea-causing pathogens, and the definition of the mechanism by which one component creates dysfunction of a critical virulence factor. The results will have an important positive impact immediately as a first step in defining and understanding the role of saliva in fighting GI disease, and long-term because they will lay the groundwork for bringing new researchers into this emerging field, and for development of saliva-based GI and respiratory therapeutics.

唾液肽抑制引起腹泻的细菌与肠道结合的能力是最新的发现但尚未得到充分表征，创新的长期目标是充分发挥唾液的潜力。作为胃肠道和呼吸道疾病的预防和治疗剂的成分。该提案的目标是确定特定唾液成分作为病原体抑制剂的功效，以及确定这些肽（Histatin-5）与菌毛结合的结构和机制产肠毒素大肠杆菌 (ETEC) 上的外部细丝这种菌毛/肽相互作用提供了抑制细菌结合的功能障碍，了解这种功能障碍的机制将导致新的发现针对 ETEC 和其他病原体的治疗方法的中心假设是增强利用。通过开发治疗方法可以实现先天免疫系统对抗疾病的作用该提案的基本原理是这项研究的完成提供了一条前进的道路。用于利用唾液成分对抗腹泻疾病的机制。 Histatin-5 的作用将允许创造出更有效的治疗方法为了实现我们的目标，我们将。追求以下两个具体目标： 1. 确定所选唾液肽抑制产肠毒素细菌结合的能力大肠杆菌 (ETEC) 作用于靶细胞； 2. 确定唾液肽 Histin-5 通过菌毛抑制细菌结合的机制。产肠毒素大肠杆菌 (ETEC) 的必需毒力因子。这些目标将通过以下方式实现：1) 对细胞培养物和原代人类进行细菌粘附研究肠道培养物（“类器官”）和 2) 使用电子进行高分辨率结构测定冷冻显微镜和冷冻断层扫描（冷冻电镜和冷冻电子断层扫描）的研究意义重大，因为它。与传统疗法不同，这将为开发针对引起腹泻的细菌的疗法开辟新途径。广泛针对参与核酸、蛋白质和细胞壁合成的酶的抗生素，在这里，我们探索宿主先天免疫系统的一个新方面：唾液成分抑制的能力这项研究的预期结果是列出了细菌对宿主的粘附。可以用作针对引起腹泻的病原体的治疗方法，并且通过以下方法定义了其机制哪一种成分会导致关键毒力因子功能障碍，结果将具有重要意义。作为定义和理解唾液在对抗胃肠道中的作用的第一步，立即产生积极影响疾病，并且是长期的，因为它们将为将新研究人员带入这一新兴领域奠定基础领域，并用于开发基于唾液的胃肠道和呼吸疗法。